• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.966-971
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• 2009

Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the $Mut^s $ Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

• Biomedical Science Letters
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• v.16 no.4
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• pp.307-310
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• 2010

Apoptosis is an important significance in the pathogenesis of cancer. Caspase 3 and p53 have been identified as important members of the apoptosis related proteins. This study was performed to define roles of caspase 3 expression and its relationship with p53 expression in endometrial cancers by immunohistochemistry. Immunoreactivity for caspase 3 was found in 13 (65.0%) out of 20 endometrial hyperplasia cases and 8 (36.4%) out of 22 endometrial cancers. Seven (87.5%) of the 8 cases with a positive caspase 3 immunoreactivity showed a positive p53 expression in 22 endometrial cancers. There were no significant associations between caspase 3 and p53 expressions. These findings suggest that caspase 3 expression might be associated with carcinogenesis of endometrial cancers. Further studies are needed to define the relationship between caspase 3 and p53 and apoptosis for examining the mechanisms of tissue-specific apoptosis related protein.

• Proceedings of the Korean Institute of Building Construction Conference
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• 2016.05a
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• pp.253-254
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• 2016

In the trend of building enlargement, high-rise and irregular shaped building, domestic application of BIM in construction phase is increasing in recent years. BIM that is based on a three-dimensional, requires a specific building expression. Model expression level required for each phase such as design, construction, and maintenance of the construction project can be different. BIM model to apply in practice is difficult because it is not clear to set up the criteria for the expression level of construction BIM model. And it has a limit that is lower productivity in construction project. Therefore, this study considers the way of the improvement and analyzes the application for representation level of construction BIM model through the survey. By analyzing of the survey results, this study will be to present a basis for an appropriate expression level setting of construction BIM model.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.51-51
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• 2003

Oct-4 is a maternally expressed octamer-binding protein encoded by the murine Oct-4 gene. It is present in unfertilized oocytes, but also in the inner cell mass and in primordial germ cells. In addition, Oct-4 is the first transcrition factor described that is specific for the blastocysts stage bovine embryos. The spatial and temporal expression patterns were further determined using Immunohistochemistry. With this technique Oct-4 protein expression is detected in the oocyte, in the blastocyst. After pregnancy Oct-4 expression is restricted ovary and placental tissue. Therefore Oct-4 is a transcription factor that is specifically expressed in cells participating in the pregnancy of Korean native cattle. These result suggest that Oct-4 localization and expression may contribute to the defects in the developmental normal seen in Korean native cattle.

• Molecules and Cells
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• v.27 no.6
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• pp.689-695
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• 2009

Chemically synthesized small interfering RNAs (siRNAs) can specifically knock-down expression of target genes via RNA interference (RNAi) pathway. To date, the length of synthetic siRNA duplex has been strictly maintained less than 30 bp, because an early study suggested that double-stranded RNAs (dsRNAs) longer than 30 bp could not trigger specific gene silencing due to the induction of non-specific antiviral interferon responses. Contrary to the current belief, here we show that synthetic dsRNA as long as 38 bp can result in specific target gene silencing without non-specific antiviral responses. Using this longer duplex structure, we have generated dsRNAs, which can simultaneously knock-down expression of two target genes (termed as dual-target siRNAs or dsiRNAs). Our results thus demonstrate the structural flexibility of gene silencing siRNAs, and provide a starting point to construct multifunctional RNA structures. The dsiRNAs could be utilized to develop a novel therapeutic gene silencing strategy against diseases with multiple gene alternations such as viral infection and cancer.

• Genomics & Informatics
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• v.14 no.1
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• pp.34-40
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• 2016

Due to advances in omics technologies, numerous genome-wide studies on human samples have been published, and most of the omics data with the associated clinical information are available in public repositories, such as Gene Expression Omnibus and ArrayExpress. While analyzing several public datasets, we observed that errors in gender information occur quite often in public datasets. When we analyzed the gender description and the methylation patterns of gender-specific probes (glucose-6-phosphate dehydrogenase [G6PD], ephrin-B1 [EFNB1], and testis specific protein, Y-linked 2 [TSPY2]) in 5,611 samples produced using Infinium 450K HumanMethylation arrays, we found that 19 samples from 7 datasets were erroneously described. We also analyzed 1,819 samples produced using the Affymetrix U133Plus2 array using several gender-specific genes (X (inactive)-specific transcript [XIST], eukaryotic translation initiation factor 1A, Y-linked [EIF1AY], and DEAD [Asp-Glu-Ala-Asp] box polypeptide 3, Y-linked [DDDX3Y]) and found that 40 samples from 3 datasets were erroneously described. We suggest that the users of public datasets should not expect that the data are error-free and, whenever possible, that they should check the consistency of the data.

• Journal of Microbiology and Biotechnology
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• v.17 no.5
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• pp.840-846
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• 2007

Human prostate-specific antigen(PSA), a 33 kDa serine protease with comprehensive homology to glandular kallikrein, is secreted from prostatic tissue into the seminal fluid and enters into the circulation. The level of PSA increases in the serum of patients with prostatic cancer and hence is widely employed as a marker of the disease status. In particular, an enzymatically active PSA that is a form cleaved at the N-terminal seven-amino-acids prosequence, APLILSR, of proPSA may play an important roll in the progression of prostate cancer. Thus, the presence of the active form would selectively discriminate the cancer from benign prostatic hyperplasia. In this study, we developed a convenient purification method for the acquisition of active PSA and proPSA. Recombinant proPSA and active PSA were expressed directly in Escherichia coli, easily and efficiently isolated from inclusion bodies, refolded, and purified. Moreover, the enzymatic activity of the recombinant active PSA was confirmed as serine protease using chromogenic chymotrypsin substrate. This purified active PSA could be further applied to scrutinize the biological or conformational characteristics of the protein and to develop specific diagnostic and/or therapeutic agents against prostate cancer.

• Biomolecules & Therapeutics
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• v.27 no.1
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• pp.78-84
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• 2019

Cell therapeutic agents for treating degenerative brain diseases using neural stem cells are actively being developed. However, few systems have been developed to monitor in real time whether the transplanted neural stem cells are actually differentiated into neurons. Therefore, it is necessary to develop a technology capable of specifically monitoring neuronal differentiation in vivo. In this study, we established a system that expresses cell membrane-targeting red fluorescent protein under control of the Synapsin promoter in order to specifically monitor differentiation from neural stem cells into neurons. In order to overcome the weak expression level of the tissue-specific promoter system, the partial 5' UTR sequence of Creb was added for efficient expression of the cell surface-specific antigen. This system was able to track functional neuronal differentiation of neural stem cells transplanted in vivo, which will help improve stem cell therapies.

• Biomedical Science Letters
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• v.22 no.4
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• pp.220-226
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• 2016

Hypertriglyceridemia induces atherosclerosis and accordingly is a major causative factor in cardiovascular diseases. Macrophages that develop into foam cells are a crucial component in the development of atherosclerosis. Monocytes can be differentiated into M1 or M2 macrophages. M1 macrophages promote inflammatory responses, whereas M2 macrophages exhibit anti-inflammatory activity. Recently, we found that triglyceride (TG)-treated THP-1 monocytes express a variety of macrophage-specific surface markers, indicating that TG treatment could trigger the differentiation of monocytes into macrophages. In this study, we investigated whether TG-induced macrophages express the M1 or the M2 macrophage phenotype. THP-1 cells were treated with various concentrations of TG for different times and the expression of M1- and M2-specific markers was evaluated by RT-PCR. We found increased expression of M1 markers (CD40, CD80, and CD86) in TG-treated THP-1 cells in a TG dose- and time-dependent manner. The expression of M2 markers (CD163, CD200R, and CD206) showed variable responses to TG treatment. Taken together, our results indicate that TG treatment triggers the differentiation of monocytes into M1 macrophages, rather than into M2 macrophages, suggesting that TG contributes to pro-inflammatory responses.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.76-76
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• 2003

Pluripotent embryonic stem cells can differentiate into beating cardiomyocytes with proper culture conditions and stimulants via embryo-like aggregates. We describe here the use of mouse embryonic stem (mES03) cells as a reproducible differentiation system for cardiomyocyte. mES03 cells growing in colonies were dissociated and allowed to re-aggregated in suspension [embryoid body (EB) formation〕. To induce cardiomyocytic differentiation, cells were exposed to 0.75% dimethyl sulfoxide (DMSO) during EB formation for 4 days and then another 4 days without DMSO (4+/4-). Thus treated EB was plated onto gelatin-coated dishes for differentiation. Spontaneously contracting colonies which appeared in approximately 4~5 days upon differentiation were mechanically dissected, enzymatically dispersed, plated onto coverslips, and then incubated for another 48~72 hrs. By RT-PCR, robust expression of cardiac myosin heavy chain $\alpha$, cardiac muscle heavy polypeptide 7 $\beta$($\beta$-MHC), cardiac transcription factor GATA4, and skeletal muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaC $h_{sm}$ ) were detected as early as 8 days after EB formation, but message of cardiac muscle-specific $\alpha$$_1$-subunit of the L-type calcium channel ($\alpha$$_1$CaCh) were reveled at a low level. In contrast, expression of myosin light chain (MLC-2V) and atrial natriuretic factor (ANF) were not detected during EB formation for 8 days. However, a strong expression of the atrial-specific ANF gene was expressed from day 8 onward, which were remained constant in EB. (cardiac specialization and terminal differentiation stage). Electrophysiological examination of spontaneously contracting cells showed ventricle-like action potential 17 days after the EB formation. This study indicates that mES03 cell-derived cardiomyocytes via 4+/4- protocol displayed biochemical and electrophysiological properties of subpopulation of cardiomyocytes.